Background: Pediatric cochlear implantation significantly improves various quality of life aspects for hearing-impaired children by influencing communication. Aim: determine the impact of instructional guidelines on knowledge and practices of mothers having children with cochlear implantation. Design: A quasi-experimental research design. Sample: A convenient sample of a total 200 of mothers and 200 children after cochlear implantation, divided equally into two groups (control& study group). Setting:  Ear, Nose, and Throat outpatient clinics at Sohag University Hospital. Tools: Tool I: Structured interview questionnaire sheet: it is composed of two parts; mothers' personal data, and children's personal data, Tool II: Mothers' knowledge about cochlear implantation (pre/post); Tool III:  Mothers' self-reported practice for their children with cochlear implantation (pre/post). Results: The majority of the mothers in the study group had a satisfactory level of knowledge and adequate level of practice toward caring for their children who have cochlear implantation post-instructional guidelines. There was a highly statistically significant difference between mothers in the study and control group regarding knowledge and self-reported practice after instructional guidelines implementation. Conclusion: The instructional guidelines improved knowledge and reported practice of mothers having children with cochlear implantation. Recommendation: Instructional guidelines for mothers about caring for their children after cochlear implantation are highly recommended.